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Cold denaturation of a protein dimer monitored at atomic resolution

Nature Chemical Biology volume 9, pages 264–270 (2013)Cite this article

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Abstract

Protein folding and unfolding are crucial for a range of biological phenomena and human diseases. Defining the structural properties of the involved transient species is therefore of prime interest. Using a combination of cold denaturation with NMR spectroscopy, we reveal detailed insight into the unfolding of the homodimeric repressor protein CylR2. Seven three-dimensional structures of CylR2 at temperatures from 25 °C to −16 °C reveal a progressive dissociation of the dimeric protein into a native-like monomeric intermediate followed by transition into a highly dynamic, partially folded state. The core of the partially folded state seems critical for biological function and misfolding.

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Figure 1: Temperature sensitivity of the CylR2 homodimer.
Figure 2: Dynamics of protein folding intermediates of CylR2.
Figure 3: Cold-induced dissociation of the native dimer.
Figure 4: The pathway of unfolding of CylR2 at atomic resolution.
Figure 5: Non-native contacts in the monomeric protein folding intermediate.
Figure 6: Three-dimensional structure of a highly dynamic, partially folded state.

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Protein Data Bank

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Biological Magnetic Resonance Data Bank

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Acknowledgements

We thank N. Rezaei-Ghaleh for useful discussions. This work was supported by the START and Ventures Programmes of Foundation for Polish Science (Fundacja na rzecz Nauki Polskiej; http://www.fnp.org.pl/) operated within the Innovative Economy Operational Programme (IE OP) 2007-2013 within European Regional Development Fund (L.J. and M.J.), the Iuventus Plus project no. IP2011 019471 from Polish Ministry of Sciences and Higher Education (M.J.) and the Cluster of Excellence and Deutsche Forschungsgemeinschaft Research Center “Nanoscale Microscopy and Molecular Physiology of the Brain” (to M.Z.). C.D.S. was supported by the Intramural Research Program of the Center for Information Technology at the US National Institutes of Health.

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Author notes

  1. Mariusz Jaremko and Łukasz Jaremko: These authors contributed equally to this work.

Authors and Affiliations

  1. Department for NMR-based Structural Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany

    Mariusz Jaremko, Łukasz Jaremko, Hai-Young Kim, Min-Kyu Cho, Karin Giller, Stefan Becker & Markus Zweckstetter

  2. Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

    Mariusz Jaremko

  3. Faculty of Chemistry, Warsaw University, Warsaw, Poland

    Łukasz Jaremko

  4. Division of Computational Bioscience, Center for Information Technology, US National Institutes of Health, Bethesda, Maryland, USA

    Charles D Schwieters

  5. German Center for Neurodegenerative Diseases (DZNE), Göttingen, Germany

    Markus Zweckstetter

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Contributions

M.J. performed structure determination, data acquisition and data analysis, and contributed to writing the paper. L.J. performed structure determination, data acquisition and data analysis, and contributed to writing the paper. H.-Y.K. performed data acquisition and analysis. M.-K.C. analyzed data, C.D.S. performed ensemble calculations, K.G. and S.B. prepared samples, and M.Z. performed data acquisition, designed and supervised the project and wrote the paper.

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Correspondence to Markus Zweckstetter.

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Jaremko, M., Jaremko, Ł., Kim, HY. et al. Cold denaturation of a protein dimer monitored at atomic resolution. Nat Chem Biol 9, 264–270 (2013). https://doi.org/10.1038/nchembio.1181

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